Janibacter Limosus Gen. Nov., Sp. Meso

INTERNATIONALJOURNAL OF SYSTEMATICBACTERIOLOGY, Apr. 1997, p. 529-534 Vol. 47, No. 2 0020-7713/97/$04.00+0 Copyright 0 1997, International Union of Microbiological Societies

Janibacter limosus gen. nov., sp. nov., a New Actinomycete with meso-Uiaminopimelic Acid in the Cell Wall KARIN MARTIN,l* PETER SCHUMA",2 FREDERICK A. RAINEY,3 BARBARA SCHUETZE,' AND INGRID GROTH' Hans-Knoll-Institut fur Naturstoff-Forschung e. V, and DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, Aussenstelle Jena, 0-07745 Jena, and DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH, 0-38124 Braunschweig, Germany

New gram-positive bacteria were isolated from 1-year-old sludge from a wastewater treatment plant. The isolates are coccoid to rod-shaped, nonmotile aerobes that form neither spores nor mycelia. They are characterized by a peptidoglycan with directly cross-linked rneso-diaminopimelic acid (type Aly), by the presence of menaquinone MK-S(H,), and by the lack of mycolic acids. The strains have complex fatty acid patterns with i-C16:oand straight-chain saturated and unsaturated fatty acids as major components. The G+C content of the DNA is 70 mol%. The results of chemotaxonomic studies and a 16s ribosomal DNA sequence comparison support our proposal to assign these bacteria to a new genus, the genus Janibacter gen. nov.; the type species is Junibucter Zirnosus sp. nov., and the type strain of J. Zirnosus is strain HKI 83 (= DSM 11140).

Both spore-forming and asporogenous actinomycetes have and starch were determined by the methods of Gledhill and Casida (10). The been screened during the last few years for useful bioactive cytochrome oxidase test was performed by using DrySlide Oxidase Test strips (Difco). The oxygen requirement was determined by using a Generbag Microaer compounds (13,39). The asporogenous organisms play impor- incubation system (bioMerieux, Marcy I'Etoile, France). The test to determine tant roles in mineralization of environmentally hazardous hydrolysis of hippurate was performed as described by Cowan and Steel (5). The chemicals, such as polycyclic aromatic hydrocarbons (22), and test to determine decomposition of gelatin was performed by using a modifica- in biotechnological production of natural products (23,26,29). tion of the method described by Lanyi (25). A UV-sterilized photographic film To isolate new actinomycetes with new biological activities, strip was immersed in phosphate buffer, which was inoculated with the test strains at a density of 6 X lo8 cells per ml (McFarland no. 2 nephelometer we investigated several samples of soil, sludge, and sewage standard). After 3 to 7 days of incubation, hydrolysis of the gelatin layer was waste. A total of 168 strains were isolated from a 1-year-old checked. Sodium chloride tolerance and growth at 28, 37, and 50°C were tested sludge sample collected from a wastewater treatment plant. on brain heart infusion (BHI) medium (Difco) and R medium. Resistance to Two of these isolates were found to be markedly different from antibiotics was tested by placing antibiotic discs (Oxoid, Hampshire, United the other isolates and from previously described genera in their Kingdom) on agar plates seeded with the test strains. Cell wall analysis. A cell wall analysis was performed as described previously chemotaxonomic and their physiological features. In this paper (14). Purified cell wall preparations were obtained by the method of Schleifer these two strains are characterized phenotypically and phylo- and Kandler (34). The amino acids and peptides of cell wall hydrolysates were genetically. Below we propose the creation of a new genus, the analyzed by thin-layer chromatography on cellulose plates by using the solvent genus Janibacter gen. nov., with one species, Janibacter limosus systems described by Schleifer and Kandler (34). The molar ratios of amino acids sp. nov., for these organisms. were determined by gas chromatography-mass spectrometry of N-heptafluoro- butyryl amino acid isobutyl esters as described previously (14). The whole-cell sugars were determined by gas chromatography of alditol acetates as described MATERIALS AND METHODS by Saddler et al. (33). The glycolate content of bacterial cells was determined by the colorimetric method of Uchida and Aida (41). Bacterial strains and cultural conditions. Strains HKI 83T (T = type strain) and HKI 84 were isolated from a 1-year-old sludge sample from the wastewater Lipid analysis. The cellular fatty acid methyl esters were determined by gas treatment plant near Jena, Thuringia, Germany, by the dilution plate technique chromatography by using the method of Stead et al. (38). Menaquinones were on plate count agar (Difco Laboratories, Detroit, Mich.). General laboratory extracted as described by Collins et a]. (4) and were analyzed by high-perfor- cultivation was performed at 28°C on solid rich medium (R medium) or in liquid mance liquid chromatography (14). Polar lipids were extracted and identified by R medium (43), which contained 1% (wtivol) Bacto Peptone (Difco), 0.5% the method of Minnikin et al. (31). The absence of mycolic acids was shown by (wt/vol) yeast extract, 0.5% (wt/vol) Casamino Acids, 0.2% (wthol) meat ex- thin-layer chromatography as described by Minnikin et al. (30). tract, 0.5% (wt/vol) malt extract, 0.2% (wtivol) glycerol. 0.1% (wt/vol) MgSO, - Analysis of DNA base composition. DNA was isolated by using a modification 7H,O, and 0.005% (wt/vol) Tween 80 (pH 7.2). To determine the cellular of the Marmur method (27). DNA was purified by treatment with proteinase K fatty acids, strains were cultivated for 24 h in liquid tryptic soy broth (Difco) at and was degraded to nucleosides with P1 nuclease and bovine intestinal mucosa 28°C. alkaline phosphatase as described by Mesbah et al. (28). The nucleosides were Morphological and physiological characteristics. Colony morphology on R separated by reversed-phase high-performance liquid chromatography as de- medium and cell morphology at different ages were determined by stereomicros- scribed by Tamaoka and Komagata (40). G+C contents were calculated from the copy and phase-contrast microscopy (Olympus, Tokyo, Japan). Nitrate reductase ratios of deoxyguanosine and thymidine. activity, urease activity, indole production, methyl red and Voges-Proskauer DNA-DNA hybridization, DNA was isolated by chromatography on hydroxy- reactions, hydrogen sulfide production, and hydrolysis of esculin and Tween 80 apatite by the procedure of Cashion et al. (1). DNA-DNA hybridization was were determined as described by Lanyi (25). Acid production from carbohy- carried out as described by De Ley et al. (6),with the modifications described by drates was determined by using the method of Hugh and Leifson (16), as Huss et al. (17), using a Gilford System model 2600 spectrophotometer equipped modified by Gledhill and Casida (10). Utilization of organic acids (2% wt/vol; with a Gilford model 2527-R thermoprogrammer and plotter. Renaturation rates sodium salts) was studied by the method of Gordon and Mihm (12). Decompo- were computed with the TRANSFER.BAS program (18). sition of adenine, hypoxanthine, xanthine, and tyrosine was determined by using 16s rDNA sequence determination and analysis. Genomic DNAs were ex- the method of Gordon et al. (11). Catalase production and hydrolysis of casein tracted from the strains investigated in this study and were used for PCR- mediated amplification of 16s ribosomal DNA (rDNA) as described by Rainey et al. (32). The purified PCR products were directly sequenced by using previously described protocols (32), and the sequence reaction mixtures were elec- * Corresponding author. Mailing address: Hans-Knoll-Institut fur trophoresed by using a model 373A automatic DNA sequencer (Applied Bio- Naturstoff-Forschung e.V., Beutenbergstraae 11, D-07745 Jena, Ger- systems, Foster City, Calif.). The 16s rDNA sequences were manually aligned many. Phone: 049-3641-656669. Fax: 049-3641-656600. E-mail: kmartin with the sequences of previously described members of the order Actinomyce- @leutra.imb-jena.de. tales. Evolutionary distances were calculated by the method of Jukes and Cantor

529 530 MARTIN ET AL. TNT. J. SYST.BACTERIOL.

following accession numbers: J. limosus DSM 11140T, Y08539; J. limosus DSM 11141, Y08540; and Microsphaera multipartita JCM 9543T, Y08541.

RESULTS Morphological and cultural characteristics. Cells of strains HKI 83T and HKI 84 are gram-positive and nonmotile and occur singly, in pairs, or occasionally in irregular clumps. While the cells of strain HKI 84 are always coccoid and vary in diameter from 0.4 to 1.1 pm (Fig. la), the cells of strain HKI 83T differ mor hologically during the growth cycle. In liquid strain HKI 83.p cultures only coccoid cell are observed, and these cells vary from 0.3 to 1.2 pm in diameter (Fig. lb). After the cells are transferred to agar slides, the coccoid cells give rise to small irregular rods (0.4 by 1.2 pm) (Fig. lc), but the cells in older cultures are coccoid. Colonies of strain HKI 83T on nutrient agar are white, opaque, and convex with glistening surfaces (Fig. 2a). The colony margins are entire. Colonies of strain HKI 84 on nutrient agar are yellow, opaque, and convex (Fig. 2b). The surfaces are matt, and the colony margins are entire. Both strains grow aerobically and in presence of 4% (wt/vol) sodium chloride in the culture medium. The optimal growth temperature is 28°C. Growth occurs at 37°C on BHI medium but not on R medium. No growth occurs under anaerobic conditions. Spore formation is not observed. Physiological characteristics. The physiological properties of isolates HKI 83T and HKI 84 are shown in Table 1.

FIG. 1. (a) Micrograph of strain HKI 84 cells grown on solid R medium at 28°C for 24 h. Bar = 10 pm. (b) Micrograph of strain HKI 83T cells grown in liquid R medium at 28°C for 24 h. Bar = 10 pm. (c) Micrograph of strain HKI 83= cells grown on an agar slide at 28°C for 12 h. Bar = 10 pm.

(20), and a phylogenetic dendrogram was constructed by using the least-squares method (7). FIG. 2. (a) Single colonies of strain HKI 83T grown on solid R medium at Nucleotide sequence accession numbers. The 16s rDNA sequences deter- 28°C for 5 days. Bar = 1 mm. (b) Single colonies of strain HKI 84 grown on solid mined in this study have been deposited in the EMBL database under the R medium at 28°C for 5 days. Bar = 1 mm. VOL. 47, 1997 JANIBACTER LIMOSUS GEN. NOV., SP. NOV. 531

TABLE 1. Morphological and biochemical characteristics of TABLE 1-Continued J. limosus HKI 83= and HKI 84 Characteristic Strain HKI 83T Strain HKI 84 Characteristic Strain HKI 83T Strain HKI 84 Chloramphenicol (30 pg) + + Color of colonies White Yellow Ciprofloxacin (5 pg) + + Morphology Coccoid, rod shaped Coccoid Erythromycin (15 pg) + + Motility Gentamicin (10 pg) + + Gram staining Kanamycin (30 pg) + + Spore formation Lincomycin (2 pg) + + Acid production from: Neomycin (30 pg) + + L-Arabinose Nitrofuran (300 pg) + + D-Cellobiose Oxacillin (5 pg) - + Dextrin Oxytetracycline (30 pg) + + D-Fructose Penicillin G (2 IU) - - D-Galactose Polymyxin B (300 IU) + + D-Glucose Rifampin (2 pg) - - Glycerol Streptomycin (10 pg) - + Inulin Sulfonamide (300 pg) - - Lactose ~ Maltose -, negative; +, positive. D-Mannitol D-Mannose D-Raffinose Chemotaxonomic characteristics. Diagnostic features that L-Rhamnose differentiate the new isolates from related genera are shown in D-Ribose Table 2. The peptidoglycan type is Aly (directly cross-linked Saccharose meso-diaminopimelic acid [meso-A,pm]), the cell wall Ala- Salicin Glu-meso-A,pm ratio for strain HKI 83T is 1.6:1.0:1.1, and D-Sorbitol Starch mycolic acids are absent. The acyl type is acetyl, the menaqui- Trehalose none is MK-8(H4), and the polar lipids are diphosphatidyl- D-Xylose glycerol, phosphatidylglycerol, and phosphatidylinositol. Diag- Utilization of nostic cell wall sugars like arabinose and galactose are not Acetate detected, but mannose, ribose, and glucose are present. The Aconitate complex fatty acid profile of strain HKI 83T is characterized by Benzoate C1711,C,7:o, and i-CI6:", as well as minor amounts of straight- Citrate chain saturated and unsaturated fatty acids (Table 3). Strain Formate HKI 84 has a complex fatty acid profile with i-C16:oand C17:1 Malate Succinate as its major components. In both strains branched-chain fatty Hippurate acids other than i-C16:ooccur only in small amounts (<3%). DL-Tartrate DNA base composition and DNA-DNA relatedness. The Decomposition of DNA base composition of both strains is 70 mol% G+C. The Adenine level of DNA-DNA relatedness between strains HKI 83T and Casein HKI 84 is 70.8%. E sc u 1 in 16s rDNA sequence analysis. Almost complete 16s rDNA Gelatin sequences of strain HKI 83T, strain HKI 84, and M. multipartita Hypoxanthine JCM 9543= comprising between 1,466 and 1,473 nucleotides Potato starch Tween 80 (>95% of the Escherichia coli sequence) were determined in Tyrosine this study. The phylogenetic dendrogram shown in Fig. 3 was Urea constructed from evolutionary distances by the least-squares Xanthine distance method (7). A total of 1,232 nucleotides present in all Nitrate reduction strains between positions 41 and 1448 (E. coli numbering) were Production of H,S used for this analysis. Figure 3 shows that strains HKI 83T and Production of indole HKI 84 fall within the radiation of the cluster comprising the Voges-Proskauer test genera Intrasporangium, Terrabacter, and Sanguibacter. The Methyl red test levels of 16s rDNA sequence similarity between organisms Oxidase test Catalase reaction belonging to this cluster range from 94.8 to 97%. The level of Growth in the presence of 16s rDNA sequence similarity between strains HKI 83T and 2% NaCl HKI 84 when all 1,473 aligned nucleotide positions were com- 4% NaCl pared was 99.4%. The levels of 16s rDNA sequence similarity 10% NaCl to the other representatives of the actinomycetes included in Growth on BHI medium at: the analysis and displayed in Fig. 3 ranged from 81.6 to 94.0%. 28°C 37°C Growth on R medium at: DISCUSSION 28°C In the course of an isolation program designed to obtain new 37°C actinomycetes that produce useful bioactive compounds, we Antibiotic susceptibility Ampicillin (10 pg) studied the chemotaxonomic and physiological properties of a large number of strains obtained from quite different sources. Continued Two of these isolates attracted our attention because of their 532 MARTIN ET AL. INT.J. SYST.BACTERIOL.

TABLE 2. Differential characteristics of the genus Janibacter and other actinomycete generaa

G+C content Diamino acid/ Mycolic Major mena- Cellular fatty Genus Polar lipids (mol%) murein type acids quinone(s) acid types

Janibacter gen. nov. 70 rneso-A,pmlAl y - MK-8(H4) DPG,PG, PI Microsphaera 67.5 rneso-A,pmlAl y - MK-8(H4) ND Brachybacteriurn" 68-72 meso-A2pmlA4y - MK-7 DPG,PG, GL DermabacteP 62 rneso-A,pmlAl y - MK-9, MK-8 PG,PL, GL Brevibacteriurn' 60-67 rneso-A,pmlAl y - MK-EI(H,), MK-7(H2) DPG,PG, GL Kineococcud 75 rneso-A,pmlAl y - MK-9(H2) DPG,PG, PGL Intrasporangiurng 68.2 ~~-A,pmlA3y - MK-8 DPG,PG, PI, PIM Terra bactefl 70-73 LL- A,pmlA3y - MK-8(H4) DPG,PI, PE,PL Nocardioidefl 66-72 ~~-A,prnlA3y - MK-8( H4) DPG,PG, PG-OH, PL Aerornicrobium" 71-73 LL- A2pm/A3y - MK-9(H4) PE, PG LuteococcuSg 66-68 LL-A2pmIA3y - MK-9(H4) DPG,PG, PI, GL Microlunatusfi 67.9 LL- A2pmlA3y - MK-9(H4) DPG,PG, PI, PL Dematophilush 57-59 rneso-A,pmlAl y - MK-8(H4) DPG,PG, PI Pseudonocardia' 79 rneso-A,pmlAl y - MK-8(H4) DPG,PG, PC, PE, PI, GL Turicelld 65-72 rneso-A,pm - MK-10, MK-11 ND Rhodococcusk 63-73 rneso-A,pmlAl y + MK-8(&) DPG,PI, PE,PIM Corynebacteriurnk 51-67 rneso-A,pmlAl y + MK-9(H2), MK-8(H2) DPG,PI, PIM Sanguibacter' 69-70 L-L~s/A~cx ND MK-9(H4) ND

a Abbreviations: LL-A,~~,LL-diaminopimelic acid; L-LYS,L-lysine; DPG, diphosphatidylglycerol; PG, phosphatidylglycerol; PE, phosphatidylethanolamine; PI, phosphatidylinositol; PIM, phosphatidylinositol mannosides; PC, phosphatidylcholine; PG-OH, phosphatidylglycerol containing 2-hydroxy fatty acids; PL, unknown phospholipid; GL, unknown glycolipid; PGL, unknown phosphoglycolipid; S, straight chain saturated; I, is0 branched; A, anteiso branched; U, monounsaturated; T, tuberculostearic acid; 20H-FA, 2-hydroxylated fatty acid; C, complex (straight chain saturated, monounsaturated, is0 branched, anteiso branched, tuberculostearic acid, and 2-hydroxylated fatty acid); ND, not determined. +, present; -, absent. Data from reference 45. Data from references 3 and 15. Data from reference 19. Data from reference 2. Data from reference 44. fi Data from reference 35. Data from reference 36. Data from reference 42. I Data from reference 9. Data from references 24 and 37. Data from reference 8. unique taxonomic characteristics. These strains were isolated celial growth and the presence of completely unsaturated only from a sludge sample that resulted from wastewater treat- menaquinone MK-8 (21). In contrast to strains HKI 83T and ment. Strains of this chemotaxonomic type were not found in HKI 84, members of the genus Sanguibacter are motile and can the other habitats examined. be distinguished by the presence of major menaquinone MK- A phylogenetic analysis based on a 16s rDNA sequence 9(H,), by having peptidoglycan type A4or with a L-Lys-Ser-D- comparison revealed that the two isolates are closely related Glu interpeptide bridge, and by a fatty acid composition whose and that their sequences are phylogenetically distinct from predominant components are anteiso-C,,,, and ClhZO(8). all actinomycete sequences available in databases. The genera While peptidoglycan type A1y is characteristic for mycolic Terrabacter, Intrasporangium, and Sanguibacter are the closest acid-containing genera (e.g., the genera Colynebacterium and phylogenetic neighbors of the new isolates (Fig. 3). The genera Rhodococcus), the following six coryneform genera are known Terrabacter and Intrasporangium differ from the new isolates in to contain meso-A,pm and to lack mycolic acids: the genera peptidoglycan type because they have type A3y peptidoglycans Brevibacterium (2),Brachybacterium (3, 15), Dermabacter (19), with LL-diaminopimelicacid, a Gly, interpeptide bridge, and a Kineococcus (44), Microsphaera (45), and Turicella (9). Mem- glycine residue bound to the or-carboxyl group of D-glutamic bers of these genera exhibit coccoid or rod-shaped morphology acid in the peptide subunit (34, 35). Differences in the fatty similar to the morphology of the strains described here. In acid patterns were also found, with the predominant fatty acid contrast to members of the genera Brachybacterium (3), Derm- i-C15:o(35) distinguishing the genera Terrabacter and Intra- abacter (19), and Kineococcus (44), members of the genus sporangium from the strains described here. Additional differ- Brevibacterium (2) and new isolates HKI 83T and HKI 84 are ences which characterize the genus Intrasporangium are my- not able to produce acid from sugars in a peptone medium.

TABLE 3. Fatty acid compositions of J. limosus HKI 83T and HKI 84

HKI 83T 0.5 0.4 1.7 0.3 3.8 0.3 17.5 0 3.9 2.4 1.3 0 0.2 20.5 29.7 1.4 0.7 2.4 8.1 HKI 84 0.7 0.2 0.7 0.1 0.9 0.3 40.8 0.3 1.3 2.6 1.0 0.2 0.1 5.0 27.6 0.3 2.9 0.7 8.5

a The abbreviations for fatty acids are illustrated by the following examples: C,,,o, hexadecanoic acid; C,,:,, octadecenoic acid; i-Cls:o, 13-methyltetradecanoic acid; ai-C,,,,, 12-methyltetradecanoic acid. i, iso; ai, anteiso. VOL. 47, 1997 JANIBACTER LIMOSUS GEN. NOV., SP. NOV. 533

the formation of mycelia and by low G+C contents (57 to 59 mol%) compared with strains HKI 83T and HKI 84 (36). The genus Pseudonocardia is characterized by the presence c of arabinose and galactose as diagnostic cell wall sugars; these Janibacter limosus HKI 83 sugars could not be detected in our isolates. The polar lipids

c Janibacter limosus HKI 84 phosphatidylcholine and phosphatidylethanolamine, which are present in members of the genus Pseudonocardia, were not found in strains HKI 83T and HKI 84 (42). On the basis of the results of our 16s rDNA sequence analysis and chemotaxonomic comparisons, we propose that the new genus Janibacter should be established for the strains described in this study. The type species of the genus Janibacter is Janibacter limosus sp. nov., which is represented by type strain HKI 83 and strain HKI 84. Dermatophilus congolensis Description of Janibacter gen. nov. Janibacter (Ja.ni.bac'ter. Micrococcus luteus L. n. Junus, a god in roman mythology, who is said to have had 7- two faces; M. L. masc. n. bacter, a rod; L. n. Janibacter, refer- Brachybacteriwn faecium ring to the changing morphology of the microorganisms). Cells are gram positive, coccoid to rod shaped, and nonmotile and Dermubacter hominis occur singly, in pairs, or occasionally in irregular clumps. No IBrevibacteriwn linem spores are formed. Not acid fast. Aerobic chemoorganotroph. Catalase is produced. Oxidase negative. The habitat of the Aemmicrobiwn erythreum strains is sludge and sewage waste. The cell wall type is Aly (directly cross-linked meso-A,pm). Mycolic acids are absent; Nocardioides albus the menaquinone is MK-8(H4). The fatty acid profile is com- MicmluMtus phosphovom plex with straight-chain saturated and monounsaturated fatty acids as the major components, and the polar lipids are diphos- Luteococcus japonicus phatidylglycerol, phosphatidylglycerol, and phosphatidylinositol. The DNA base composition is 70 mol% G+C. The type Sporichthya polymorpha species is Janibacter limosus. Geodermatophilus obscurus Description of Janibacter limosus sp. nov. Janibacter limosus (1i.mo'sus. L. adj. limosus, muddy, pertaining to sludge, the Micmsphaera muitipanita natural habitat of the species). Cells are gram-positive and nonmotile and occur singly, in pairs, or occasionally in irregu- Rhodococcus rhodochmus lar clumps. In liquid cultures cells are coccoid (diameter, 0.3 to Nocardia astemides 1.2 km). On solid medium strains vary in cell morphology. The cells of some strains are short rods (0.4 by 1.4 km), which Lenrzea albidocapillata change shape and become coccoid in older cultures. Colonies on nutrient agar are white or yellow, opaque, and convex; the Pseudonocardia sarumea colony surfaces are glistening or matt; and the colony margins are entire. limosus grows aerobically and tolerates 4% (wt/ Srreptomyces albidofivus J. vol) NaCl in the medium. The optimal growth temperature is Atopobium minutum 28°C; growth occurs at 37°C on BHI medium but not on R medium. No growth occurs under anaerobic conditions. Cata- lase, indole, and H,S are produced; urease and oxidase are not FIG. 3. Phylogenetic dendrogram based on the results of a 16s rDNA se- produced. Gelatin, starch, Tween 80, and casein are hydro- quence comparison. Bar = 5 nucleotide substitutions per 100 nucleotides. lyzed. Citrate, formate, malate, and tyrosine are decomposed; aconitate, benzoate, succinate, tartrate, hippurate, esculin, adenine, xanthine, and hypoxanthine not decomposed. Nitrate is reduced to nitrite. Voges-Proskauer and methyl red reactions Strains HKI 83T and HKI 84 can be easily distinguished from are negative. No acid is produced from L-arabinose, cellobiose, members of the genera Brevibacterium, Brachybacten'um, dextrin, fructose, galactose, glucose, glycerol, m-inositol, inu- Dermabacter, Kineococcus, and Tun'cella on the basis of the lin, lactose, maltose, mannitol, mannose, raffinose, L-rham- presence of menaquinone MK-8(H4) and differences in the nose, ribose, salicin, D-sorbitol, sucrose, potato starch, treha- fatty acid profiles. Compared with strains HKI 83T and HKI lose, and D-xylose. Has the chemotaxonomic characteristics of 84, members of the genera Brevibacten'um and Dermabacter are the genus. The DNA base composition is 70 mol% G+C. The characterized by lower DNA G+C contents and members of type strain is strain HKI 83 (= DSM 11140). the genus Kineococcus are characterized by higher DNA G+C contents (Table 2). Strains HKI 83T and HKI 84 have the same peptidoglycan type, (type Aly) and the same menaquinone ACKNOWLEDGMENTS [menaquinone MK-8(H4)] as members of the genera Micro- We thank Christiane Weigel, Carmen Schult, Karin HoOrich, and sphaera, Dermatophilus, and Pseudonocardia but are phyloge- Renate Schumann for excellent technical assistance. Jutta Burghardt is netically unrelated to these taxa (Fig. 3). M. multipartita also thanked for carrying out DNA-DNA reassociation experiments. differs in its predominant fatty acids (iso-C,,,,, iso-CIs:o, and This work was supported by grant 0310591A from the Bundesmin- C18:l) and in having a lower G+C content (67.5 mol%). Rep- isterium fur Bildung, Wissenschaft, Forschung und Technologie, Ger- resentatives of the genus Dermatophilus are characterized by many. 534 MARTIN ET AL. Im. J. SYST.BACTERIOL.

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